945;-glucanase Inhibits Conidial Agglutination of Aspergillus fumigatus and Promote Inflammatory Cytokine Production from THP

Ref ID: 19611

Author:

S Ide1*, Y Imamura1, S Yano2, K Takeda1, M Yoshida1, K Hirano1, T Takazono1, K Kosai1,3,
Y Morinaga4, S Nakamura1, S Kurihara1, M Tsukamoto1,3, K Izumikawa1,3, K Yanagihara4,
T Tashiro1, S Kohno1

Author address:

1Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences,
Nagasaki, Japan
2Biochemical Engineering, Yamagata University Graduate School of Science and Engineering, Yamagata,
Japan
3Infection Control and E

Full conference title:

6th Advances Against Aspergillosis 2014

Abstract:

Purpose:
α -glucan is one of the main component of cell wall of Aspergillus fumigatus swollen conidia,
hyphae and extracellular matrix and it covers β -glucan. We examined the influence of α -glucan-lytic
enzyme α -glucanase (mutanase) from Bacillus circulans for growth of A. fumigatus and production
of inflammatory cytokines from THP-1 stimulated with A. fumigatus.
Methods:
First, we evaluated expression of cell wall α -glucan and β -glucan sequentially using immunofluorescent
stain with α -glucanase. Secondly, A. fumigatus were cultured in shaking incubator with α -glucanase
for 3 and 24 hours and assessed agglutination of conidia. Thirdly, we assessed conidial adhesion
to human lung epithelial cell H292 with α -glucanase. Finally, human monocytic cell THP-1 were
stimulated with LPS and germinating conidia of A. fumigatus preincubated for 12 hours with
α -glucanase. We assessed production of inflammatory cytokines.
Results:
Fluorescent intensity of cell wall α -glucan and β -glucan were observed after swollen conidia,
although they were not expressed on resting conidia. The intensity of α -glucan reduced with
α -glucanase. Conidial agglutination in shaking culture and conidial adhesion to H292 cell layer
were inhibited by α -glucanase. THP-1 produced inflammatory cytokine interleukin-8 (IL-8) and
tumor necrosis factor-alpha (TNF-α ) stimulated with germinating conidia. Additionally, α -glucanase
induced production of IL-8 and TNF-α from THP-1 stimulated by Aspergillus conidia, although it
had no influence on the production by LPS.
Conclusions:
On Aspergillus cell wall, β -glucan, which is recognized from host immune cells by Dectin-1, is
covered with α -glucan in growing phase. Previous reports suggest that α -glucan is not essential
for growth of Aspergillus. Our results suggest that α -glucanase inhibits initial agglutination and
adhesion of conidia and promotes production of inflammatory cytokines lysing α -glucan covering
β -glucan in growing phase.

Abstract Number: 136

Conference Year: 2014

Link to conference website: http://www.AAA2014.org

New link: NULL

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